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CN-122012601-A - Gene for regulating and controlling wheat flower formation transformation and agronomic characters and application thereof

CN122012601ACN 122012601 ACN122012601 ACN 122012601ACN-122012601-A

Abstract

The application discloses a gene for regulating and controlling wheat flower formation transformation and agronomic characters and application thereof, and belongs to the technical field of genetic engineering. The application discloses a gene related to agronomic traits such as wheat flowering transformation, adventitious small flower spike, adventitious root and the like and application thereof, and belongs to the technical field of genetic engineering. The application takes VRN3 genes on Chinese spring 7A, 7B and 7D chromosomes as reference genome, and designs knockout targets in the conserved regions. The expression of the gene is regulated and controlled through a wheat stable genetic transformation experiment so as to verify the effect of the gene on the transformation of wheat from vegetative growth to reproductive growth and other agronomic traits. The application of the VRN3 gene provides a theoretical basis for cultivating multi-floret variety materials and asexual reproduction wheat materials. In addition, by utilizing VRN3 gene, sterile line of hybrid seed production system can be created in future, thus providing new way for establishing high-efficiency hybrid seed production system of wheat.

Inventors

  • FU DAOLIN
  • GONG TINGTING
  • WANG WENQIANG
  • Hao Qunqun
  • Zou Chunhao
  • ZHANG HUIFEI
  • LIU FANG
  • GONG SANG

Assignees

  • 泉脉农业科技有限公司

Dates

Publication Date
20260512
Application Date
20260402

Claims (10)

  1. 1. Use of the wheat VRN3 gene in any one of the following (1) - (3): (1) Producing adventitious spikes or complex spikes, resulting in a change in the structure of the spike; (2) Generating adventitious roots at the stalk nodes; (3) Cultivating wheat varieties with multiple flowers and multiple spikes; (4) Cultivating perennial or apomictic plant material; The wheat VRN3 gene is expressed by TraesCS7A02G115400.1, traesCS7B02G0130.1 or TraesCS7D02G111600.1.
  2. 2. The use according to claim 1, wherein the adventitious roots produced at the stalk node are viable after transplanting and develop into new independent plants.
  3. 3. The use according to claim 1, characterized in that by knocking out the VRN3 gene, the ear structure of the wheat plant is altered and/or adventitious roots are produced at the stalk node.
  4. 4. The use of claim 1, wherein said knocking out the VRN3 gene comprises frameshift mutating said gene or creating a loss of function mutation on at least one allele.
  5. 5. Use of a protein encoded by the wheat VRN3 gene of claim 1 in (1) or (2) as follows: (1) Producing adventitious spikes or complex spikes, resulting in a change in the structure of the spike; (2) Adventitious roots are produced at the stalk nodes.
  6. 6. The use according to claim 5, characterized in that the ear structure of the wheat plant is altered and/or adventitious roots are produced at the stalk node by reducing the expression of the protein encoded by the wheat VRN3 gene; or by inactivating the protein encoded by the wheat VRN3 gene, altering the ear structure of the wheat plant and/or producing adventitious roots at the stalk node.
  7. 7. A method of creating plant material capable of asexual reproduction or having perennial habit, comprising the steps of: (1) Constructing a CRISPR-Cas9 gene editing vector aiming at a wheat VRN3 gene, introducing the CRISPR-Cas9 gene editing vector into a plant receptor cell, and regenerating and obtaining a transgenic plant; (2) Positive plants were selected that produced an adventitious root phenotype at the stalk node during the vegetative growth stage.
  8. 8. The method of claim 7, wherein the method is used to create asexual wheat, perennial wheat, pasture wheat, or wheat material for greening.
  9. 9. A method for asexual reproduction of wheat, which is characterized by comprising the steps of obtaining a positive wheat plant which generates an adventitious root phenotype at a stalk node in a vegetative growth stage according to the method described in claim 7, cutting a tillering with an adventitious root of a stalk node, and transplanting and culturing the tillering to obtain a progeny plant with the same genetic background through the asexual reproduction method.
  10. 10. A gene-edited wheat plant obtained by knocking out or inactivating a wheat VRN3 gene, characterized in that said plant has one or more of the following traits (a) to (e): (a) Producing adventitious spikes or complex spikes; (b) Generating adventitious roots at the stalk nodes; (c) Delayed flowering or long-term vegetative growth; (d) The plant height is reduced; (e) Pollen fertility is reduced or sterile.

Description

Gene for regulating and controlling wheat flower formation transformation and agronomic characters and application thereof Technical Field The invention belongs to the technical field of genetic engineering, and particularly relates to a gene for regulating and controlling wheat flower formation transformation and agronomic traits and application thereof. Background The process of wheat from vegetative growth to reproductive growth is the result of the co-regulation of genes and the environment. Flowering time plays an important role in plant propagation offspring. Among the three types of genes (vernalization gene, photoperiod gene and premature gene), the vernalization gene has important roles in controlling the whole growth cycle length of wheat to be 70% -75%, participating in the related processes of plant heading, flowering and the like. There are four major genes known at present for controlling the vernalization process of wheat, VRN1, VRN2, VRN3 and VRN4. The time to reach the two-edge period of the wheat material carrying different near isogenic lines of the vernalization gene has obvious difference, which shows that different genotypes of the vernalization gene have a certain influence on the flowering time of the wheat, regulate the heading period and winter-spring property of the wheat and control the flowering time of the wheat. The research of the interaction between vernalization genes has important significance for breeding, introduction, cultivation, production and utilization. Salina E et al found from a study of heading-related genes of 95 common wheat varieties that VRN-B1 and VRN-B3 contributed most to the genetic diversity of wheat, and wheat varieties containing different alleles could be potential sources for creating natural variation of suitable materials in heading period (Berezhnaya et al., 2021). Chen et al found that vernalization genes and their copy number variations had an important effect on plant flowering and maturation, which had an important significance for cultivating early maturing varieties, avoiding the effect of dry hot air on wheat yield reduction (Chen et al, 2013). Studies have shown that the VRN-A3 allele is the primary cause of flowering that cultivated EMMER WHEAT can be achieved under low latitude, short sun conditions (Nishimura et al 2021). The wheat seed production and selection system is a key step in the wheat breeding process. The SPT technology is the earliest proposed hybrid seed production and selection system, and the technology realizes the screening of offspring seeds by adding fluorescent protein genes to obtain target character materials. In agricultural production, the use of gene editing crops may have potential safety hazards, require approval by regulatory authorities and have complicated procedures, however, the hidden safety problem in gene editing can be solved by obtaining wheat materials which cannot be used for seed production through gene editing. The current SPT technology mainly relies on the utilization and excavation of sterile genes to realize the maintenance and propagation of sterile lines, and compared with cytoplasmic male sterility and nuclear cytoplasmic interactive male sterility, the utilization and development of nuclear sterile genes with stable inheritance can solve the problems of inbred fecundity of cytoplasmic female parent genetic F 1, single cytoplasm of hybrid seeds, low purity yield and the like. Like many annual crops, wheat cultivated today belongs to annual or perennial herbaceous plants. Unlike perennial crops, which have developed root systems, they can prevent water and soil loss, better absorb and utilize soil nutrients, and annual crops have lower moisture and nitrogen fertilizer utilization rates (Glover et al, 2010; randall & Mulla, 2001). Perennial growth habit is quantitative character controlled by multiple genes, and genetic mechanism is complex. Zhai et al found three MADS-box genes FLOWERING LOCUS C (FLC), FLOWERING LOCUS M (FLM) and MADS AFFECTING FLOWERING (MAF) in Arabidopsis that determine plant perennial, and the transformation of materials into perennial was achieved by introducing any one of these genes into annual materials (Zhai et al 2024). Madrid et al have studied annual and perennial materials of Arabidopsis by genetic, transgenic and genomic means and found that AmMAR a is an important flowering inhibitor, indirectly leading to perennial and annual life habits (Madrid et al 2021). The perennial wheat not only can provide precious resources for variety cultivation and quality improvement, but also can be used for grassland ecological management, cultivation of pasture materials, increase of green land coverage area, and become an important direction for future wheat research. The control of the excavation and application of the excellent character genes of wheat has important significance for cultivating new variety materials of wheat, expanding the application path of wheat and p